Browsing by Author "Smith, Savanna"
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Item Brain mitochondrial dysfunction in postpartum preeclamptic rodents(2023) Jones, Kylie; Smith, Jonna; Smith, Savanna; Castillo, Angie; McCafferty, Adair; Wiemann, Natalia; Owen, Malissa; Srivastava, Prakriti; Cunningham, MarkPurpose: Pre-eclampsia (PE), new-onset hypertension during pregnancy, impacts 3-8% of all births in the USA yearly and causes significant neurological damage to the mother during and after pregnancy. Studies show postpartum PE women to have increased risks of hypertension (HTN) and cerebrovascular dysfunction (CVD). Although the cause of HTN and cerebral damage is unknown, mitochondrial dysfunction (mtDys) may play a role. MtDys includes reduced mitochondria-specific antioxidants, raised mitochondrial reactive-oxygen species, changes in mitochondrial fission and fusion proteins, and reduced efficiency of the electron transport chain (ETC). Previous studies in our lab indicate associations between cardiac mtDys and the reduced uterine perfusion pressure (RUPP) rat model of PE with HTN at 10 weeks postpartum (PMID: 34727994). However, cerebral mtDys has not been examined in RUPP rats postpartum. This study aims to examine cerebral mitochondrial functional proteins in hypertensive RUPP postpartum rats at six weeks. We hypothesize that RUPP postpartum rats will have lower amounts of cerebral mitochondrial functional proteins compared to control (CON) postpartum rats. Methods: We divided pregnant Sprague Dawley rats into two groups: CON normal pregnant (NP, n = 4) and RUPP (n = 4). Then, the RUPP surgery was performed on gestational day 14. Pregnant rats gave birth naturally and weaned for three weeks. Six weeks after giving birth, rats were euthanized for brain collections to measure functional proteins via Western Blot analysis, including ETC complexes (Complexes I-V), fusion proteins (OPA-1 and MFN-2), fission protein (DRP-1), and mitochondria-specific antioxidant (MnSOD). Results: In the brain, RUPP postpartum rats have significantly reduced Complex I proteins compared to NP postpartum rats (91 ± 2.27 vs. 100 ± 2.45 IU/protein/CON %, p < 0.05) with slight decreases in Complexes II (93 ± 4.14 % vs. 100 ± 7.57 IU/protein/CON %, ns), III (91 ± 3.18 vs. 100 ± 6.11 IU/protein/CON %, ns), IV (86 ± 11.25 vs. 100 ± 7.95 IU/protein/CON %, ns), and V (92 ± 3.99 vs. 100 ± 6.33 IU/protein/CON %, ns). RUPP and NP postpartum rats have no significant differences in fusion proteins OPA-1 (102 ± 2.56 vs. 100 ± 2.02 IU/protein/CON %, ns) and MFN-2 (106 ± 18.25 vs. 100 ± 14.35 IU/protein/CON %, ns). Fission protein DRP-1 has an increase in RUPP postpartum rats compared to NP postpartum rats (111 ± 6.92 vs. 100 ± 4.55 IU/protein/CON %, ns). RUPP postpartum rats have significantly decreased MnSOD in comparison to NP postpartum rats (89 ± 2.00 vs. 100 ± 2.45 IU/protein/CON %, p < 0.05). Conclusion: RUPP postpartum rats have cerebral mtDys indicated by decreased ETC complexes, especially Complex I. RUPP postpartum rat brains have reduced MnSOD, which suggests elevated mitochondrial oxidative stress. Furthermore, raised mitochondrial fission in the brain supports the presence of mitochondrial damage and mtDys. Future studies will examine the role of cerebral mtDys in causing HTN and CVD in RUPP postpartum rats. This study is clinically relevant because our findings provide a possible mechanism for the pathophysiology of CVD in postpartum PE women and novel targets for cerebral mitochondrial therapy.Item Changes in Blood Pressure and Abundance of Kidney Sodium Channels in Postpartum Preeclamptic Rats(2023) McCafferty, Adair; Smith, Jonna; Smith, Savanna; Jones, Kylie; Castillo, Angie; Wiemann, Natalia; Owen, Malissa; Srivastava, Riti; Cunningham, MarkPurpose: Postpartum (PP) preeclamptic (PE) women have an increased risk of developing hypertension (HTN) and chronic kidney disease. However, the mechanisms of these diseases in PP PE women are not fully understood. Results from our previous studies show 10-week PP PE rats have HTN (PMID: 34727994). One factor that could cause elevated blood pressure is an increase in sodium transport in the kidneys of PP PE rats. Our current study will examine the BP as well as the amount of Sodium Potassium Chloride Cotransporters (NKCC), Sodium Hydrogen Exchanger-1 (NHE1) and a-subunit of the Epithelial Sodium Channel (ENaC) within the kidney of PE rats 6 weeks postpartum (PP6). We hypothesize that PP6 PE rats will have an increase in BP as well as increased amounts of NKCC, NHE1 and a-ENaC proteins. Methods: Pregnant Sprague Dawley rats were divided into 2 group: control (CON) normal pregnant rats and PE rats (derived from the surgically induced placental ischemic model of PE). All rats gave birth and were weaned for 3 weeks. At PP6, BP was measured via carotid catheterization. Kidney cortex (KC) and medulla (KM) tissue were collected to measure for NKCC, NHE1 and a-ENaC amounts via Western Blots. Results: BP was significantly elevated in PP6 vs CON rats (128 ± 6 vs 106 ± 4mmHg, p<0.05). The amount of NKCC channels within the KC was significantly increased in PP6 vs CON rats (107.82 ± 5.64 vs 100 ± 1.48 IU/Protein/CON%, p<0.05). The amount of a-ENaC proteins within the KC was also increased in PP6 vs CON rats (123.4 ± 4.67 vs 116.5 ± 33.03 IU/Protein/CON%, p=0.06). NHE1 channels within the KC were slightly decreased in PP6 vs CON rats (77.31 ± 15.28 vs 100 ± 16.61 IU/Protein/CON%, N.s). No changes were observed in a-ENaC proteins within the KM in PP6 vs CON rats. Conclusion: PP6 PE rats have HTN as well as increased NKCC and a-ENaC sodium transport proteins in the kidney cortex. These results confirm our hypothesis that increased sodium transporters in the kidney, which could elevate sodium and water reabsorption, correlates with an increase in blood pressure. This study is clinically relevant because it informs clinicians on the pathophysiology of HTN and renal disease/dysfunction in PP PE women. It also highlights novel approaches to providing potential therapies to manage blood pressure in PP PE women. Future studies will investigate the cause-and-effect relationship between blood pressure and sodium handling in the kidney of PP PE rats.Item Changes in cardiac oxidative stress, nitric oxide bioavailability, mitochondrial function, and blood pressure in postpartum preeclamptic rats(2023) Owen, Malissa; Smith, Jonna; Smith, Savanna; Jones, Kylie; Castillo, Angie; Wiemann, Natalia; McCafferty, Adair; Srivastava, Prakriti; Cunningham, MarkBackground: Preeclampsia (PE) is characterized by new onset hypertension (HTN) during pregnancy that usually occurs in the third trimester and is associated with decreased nitric oxide (NO) bioavailability, increased oxidative stress (OS), and mitochondrial dysfunction. Postpartum (PP) PE women have an increased risk of developing HTN and cardiovascular diseases (CVD) later in life. The timing and mechanisms of this rise in blood pressure (BP) and cardiovascular dysfunction in PP PE women are unknown. Previous studies in our lab indicate that PP PE rats have HTN, increased OS, and cardiac mitochondrial dysfunction at 10 weeks (PP10) (PMID: 34727994). Our current study examines the relationship between BP and cardiac NO bioavailability, OS, and mitochondrial dysfunction at 6 weeks (PP6), an earlier time point from our previous study. We hypothesize that PP6 PE rats will have HTN, increased cardiac OS, decreased cardiac NO bioavailability, and mitochondrial dysfunction. Methods: Pregnant Sprague Dawley rats were divided into 2 groups: normal pregnant rats (NP) and PE rats, derived from the surgically induced reduced uterine perfusion pressure model of PE (RUPP). All rats gave birth, and their offspring were weaned for 3 weeks. At PP6, BP was measured via carotid catheterization and heart tissues were collected to measure heat shock protein (HSP-1) (a measure of OS), copper zinc superoxide dismutase (CuZnSOD) (an antioxidant), manganese SOD (MnSOD) (a mitochondrial specific antioxidant), endothelial NOS (eNOS) (a measure of NO producing enzyme), and electron transport chain (ETC) proteins (a measure of mitochondrial function). These quantities were obtained through Western blots. Results: BP was significantly elevated in PP RUPP vs PP NP rats (128±6 vs 106±4mmHg, p < 0.05). HSP-1 was significantly decreased in PP RUPP vs PP NP rats (88±1.51 vs 100±4.05IU/Protein/CON%, p < 0.05). CuZnSOD showed no change between the two groups while MnSOD was drastically increased in PP RUPP vs PP NP (123±2.91 vs 100±5.30IU/Protein/CON%, p < 0.05). eNOS and ETC proteins were unchanged between PP RUPP and PP NP rats. Conclusion: Contrary to our hypothesis, PP6 PE rats have HTN with no increase in cardiac OS (due to decreased HSP-1 and increased in MnSOD amount) or decrease in cardiac NO bioavailability and mitochondrial dysfunction. These observations are different from our 10-week PP PE rats and may suggest that the heart is protected at 6 weeks PP despite the increase in blood pressure. Future studies will focus on the time frame in which cardiac dysfunction occurs in PP PE rats. Additionally, studies will explore the antioxidant, NO, and mitochondrial pathways along with other cellular mechanisms that may prevent the heart from damage and/or dysfunction after a PE pregnancy. This study is clinically relevant because it will inform clinicians on the mechanisms of HTN and cardiac dysfunction in women following a PE pregnancy as well as provide insights on therapies that could be used to prevent the development of CVDs later in life for PP PE women.Item Changes in cerebral inflammation and blood pressure in postpartum preeclamptic rats(2023) Wiemann, Natalia; Smith, Jonna; Smith, Savanna; Jones, Kylie; Castillo, Angie; Cromartie, Whitney; Owen, Malissa; McCafferty, Adair; Srivastava, Prakriti; Cunningham, MarkIntroduction: Postpartum (PP) preeclamptic (PE) women have an increased risk of developing hypertension (HTN) and cerebrovascular diseases later in life. Studies show that women who experience preeclampsia go on to develop HTN 7-8 years earlier than women with normal pregnancies, increasing their risk for developing cerebrovascular diseases such as stroke. While the timing and mechanisms contributing to a rise in blood pressure (BP) and cerebrovascular dysfunction in postpartum preeclamptic women are unknown, they are hypothesized to be influenced by inflammation. Previous studies in our lab indicate postpartum PE rats at 10 weeks have HTN and increased inflammation (PMID: 34727994). Our current study will examine BP and inflammation in postpartum PE rats at an earlier time point, 6 weeks (PP6), to determine the relationship between cerebral inflammation and the pathophysiology of HTN. We hypothesize that postpartum PE rats will have an increase in BP and cerebral inflammation 6 weeks after delivery. Methods: Pregnant Sprague Dawley rats were divided into 2 groups: normal pregnant (NP) rats, and preeclamptic (RUPP) rats. Placental ischemia was surgically induced in the preeclamptic group via the reduced uterine perfusion pressure (RUPP) model. All rats gave birth naturally and were weaned for 3 weeks. At PP6, BP was measured via carotid catheterization, and brain tissue was collected to measure pro-inflammatory (TNF-α and IL-17) and anti-inflammatory (IL-4 and IL-10) factors via colorimetric assays and ELISAs. Results: Blood pressure was elevated in RUPP PP vs NP PP (128±6 vs 106±4mmHg, p<0.05). Cerebral TNF-α drastically increased by ~2.4 fold in RUPP PP vs NP PP (2576±445.6 vs 1058±212.5pg/mL, ns). Cerebral IL-17(331.2±41.1 vs 297.6±48.6pg/mL, ns) and IL-4 (178.4±23.4 vs 154.8±14pg/mL, ns) also increased in RUPP PP vs NP PP. Cerebral IL-10 (103.9±21.4 vs 147.6±11.3pg/mL, ns) was decreased in RUPP PP vs NP PP rats. Conclusion: PP6 preeclamptic rats have HTN and increased cerebral inflammation. It is yet to be determined whether cerebral inflammatory markers are the cause or consequence of HTN in PP6 PE rats. Future studies will explore the sequence of HTN and cerebral inflammation in postpartum PE rats and determine how brain inflammation contributes to HTN and cerebral damage. We will also target specific areas of the brain that modulate autonomic function and BP. This study is clinically relevant because it will inform providers on the pathophysiology of HTN and/or cerebral damage in women after a PE pregnancy. Our findings suggest that the use of inflammatory therapeutic targets improves HTN and cerebrovascular dysfunction in postpartum PE women.Item Changes in Cerebral Mitochondrial Function in Postpartum Dams Exposed to a Low-resource Environment during Weaning(2024-03-21) Jones, Kylie; Smith, Savanna; Smith, Jonna; Castillo, Angie; Powell, Madison; Cunningham, MarkBackground: Postpartum depression is a serious mental-health condition, affecting over 14% of all mothers in the U.S. Poverty, a lack of educational and economic resources, is a major determinant of adult mental health. In the U.S., poverty impacts 11% of adults, with a greater incidence in women of childbearing age (roughly 16%) and postpartum women (roughly 14%). While pregnancy and poverty separately increase risks of developing depression, few studies have combined these conditions to determine their role in the development of depression. Further, no studies have explored mechanisms contributing to depression following pregnancy and poverty. However, cerebral mitochondrial dysfunction (C-mtDys) may be one mechanism. The aim of this study is to examine C-mtDys in postpartum (PP) dams exposed to the limited-bedding-nesting (LBN) model in rats, which reduces their nesting material to simulate a low-resource environment. We hypothesize that PP dams in the LBN model will exhibit C-mtDys and elevated oxidative stress. Methods. Pregnant Sprague-Dawley rats gave birth naturally and were divided randomly into LBN (n =2) or control (n = 2) groups. LBN dams were exposed to the LBN model from PD 2 through PD 9. To validate the success of the LBN model, entropy scores, which are a measure of behavior unpredictability, were recorded for each dam. At 17 weeks PP, equivalent to 8 years PP in humans, brains were collected and used to isolate mitochondria through differential centrifugation. Mitochondrial function was evaluated via respiration (mtRes) using respiratory states. Oxidative stress in the whole brain was examined through H2O2 and total antioxidant capacity biochemical assays. Results. LBN PP dams displayed higher entropy scores (1.10 ± 0.04 v. 0.79 ± 0.04, p < 0.05), as expected based on prior literature, serving as validation of the model. At 17 weeks, PP LBN dams had reduced mtRes in all states, including the basal state (176.09 ± 5.90 v. 360.70 ± 7.73 pmol/s/mg, ns), State 2 (1044.72 ± 14.00 v. 1703.00 ± 18.10 pmol/s/mg, ns), State 3 (3843.27 ± 31.86 v. 6705.72 ± 37.54 pmol/s/mg, ns) and State 4 (776.83 ± 13.65 v. 1533.54 ± 16.46 pmol/s/mg, ns). PP LBN dams exhibited reduced total antioxidant capacity by roughly 20% (54.10 ± 1.24 v. 63.04 ± 0.99 mM Trolox) and elevated H2O2 by roughly 50% (2.12 ± 0.52 v. 1.35 ± 0.28 nM/mg, ns). Summary. In summary, preliminary data shows C-mtDys via reduced mtRes in PP dams exposed to an impoverished environment during weaning. Furthermore, this study suggests that decreased mtRes could contribute to increased oxidative stress in the brain. Elevated oxidative stress may cause damage at the cellular and circuitry levels in the brain that could facilitate the development of depression later in life. Future studies will further examine C-mtDys, oxidative stress, and depressive behaviors in PP dams exposed to LBN. This study is significant because it identifies C-mtDys as a possible mechanism causing depression after exposure to both pregnancy and poverty.Item Early Life Sex Differences and Alterations in Mitochondrial Function in IUGR Offspring after Weaning may Contribute to Adult Sex Differences in Cerebrovascular Dysfunction and Hypertension in Rodents(2024-03-21) Hart, Savannah; Smith, Savanna; Smith, Jonna; Jones, Kylie; Castillo, Angie; Davis, Zandria; Springfield, Alex; Rollings, Alyssa; Narra, Sreeram; Cunningham, MarkBackground: Preeclampsia is a hypertensive pregnancy disorder that usually occurs in the third trimester. Preeclampsia, due to placental ischemia, decreases nutrient and oxygen delivery to the fetus, causing intrauterine growth restriction (IUGR). IUGR increases the risk for chronic conditions such as cerebrovascular dysfunction (CVD) and hypertension (HTN). Human and animal studies show sex differences in CVD and HTN development in IUGR offspring, with males exhibiting a higher prevalence of both. Preliminary data from our lab showed that 17-week-old IUGR male rodents developed CVD, HTN and mitochondrial dysfunction (mtDYS), while IUGR female rodents only developed CVD. The reason for these sex differences is unknown but may be attributed to mtDYS. Although mitochondrial differences appear at 17 weeks, earlier mitochondrial function is unknown. This study investigates changes in mitochondrial function in IUGR offspring after weaning. We hypothesize that mtDYS is elevated in3-week-old IUGR rodent offspring, with greater dysfunction in IUGR males. Methods: Pregnant Sprague Dawley rats were divided into two groups: normal pregnant (NP) and preeclamptic pregnant rats, which underwent reduced uterine perfusion pressure (RUPP) surgery on gestational day 14. RUPP dams gave birth to IUGR offspring, and NP dams gave birth to control (CON) offspring. After 3 weeks of weaning, offspring were separated by sex and dam pregnancy status. Brains were collected from the following groups: IUGR males (n=6), IUGR females (n=6), CON males (n=4), and CON females (n=6) to measure mitochondrial function via respiration, electron transport chain (ETC) protein amounts, and mitochondrial dynamics of fission (DRP-1) and fusion (MFN-1) proteins. Mitochondrial respiration was assessed using the Oroboros Oxygraph O2K. Protein amounts of ETC complexes (I-V), DRP-1, and MFN-1 were quantified using Western blots. Results: 3-week-oldIUGR females had increased cerebral mitochondrial respiration suggested by State 3 (490.41 ± 49.85 vs 257.32 ± 69.76pmol O2/sec/mg; p=0.02) and increased protein amounts of ETC Complex I (135.91 ± 9.27 vs 107.77 ± 4.23IU/Protein/CON%; p=0.02) and Complex III (141.76 ± 13.99 vs 110.43 ± 7.73IU/Protein/CON%; p=0.07). MFN-1 protein amounts (133.42 ± 18.75 vs 84.67 ± 6.89IU/Protein/CON%; p=0.03) and DRP-1 protein amounts (111.31 ± 1.89 vs 95.24 ± 2.14IU/Protein/CON%; p<0.0003) were increased in IUGR compared to CON females. Conversely, 3-week-old IUGR males showed decreased cerebral mitochondrial respiration in Basal state (21.48±5.49 vs 55.79 ± 6.96pmol O2/sec/mg; p=0.03) and State 2 (146.22 ± 25.55 vs 224.70 ± 23.13pmol O2/sec/mg; p=0.13). DRP-1 protein amounts were decreased in IUGR males (85.72 ± 3.07 vs 110.18 ± 1.82IU/Protein/CON%; p=0.02), with no changes in mitochondrial ETC complexes or MFN-1 protein amounts compared to CON males. Conclusion: Early on, IUGR females show mitochondrial function, while IUGR males display mtDYS. Furthermore, the mtDYS in IUGR males at 3 weeks may contribute to HTN development observed in IUGR males and not IUGR females at 17 weeks of age. Future studies are warranted to investigate mtDYS and HTN development in IUGR males and possible protective mechanisms in IUGR females. This study highlights sex differences in cerebral mitochondrial function in prepubescent IUGR offspring, offering insight into the pathophysiology of HTN and CVD development in adulthood, along with suggestions for novel therapeutic targets to prevent HTN and CVD in adult IUGR offspring.Item Elevated Renal Oxidative Stress and Na+ Transporters are Associated with Hypertension in Postpartum Preeclamptic Rats(2024-03-21) Smith, Savanna; Castillo, Angie; Jones, Kylie; Smith, Jonna; Hart, Savannah; Powell, Madison; Cunningham, MarkApproximately 5-10% of US pregnancies result in preeclampsia (PE). PE is characterized by new onset hypertension (HTN) during pregnancy and is usually accompanied by end-organ damage, especially in the kidneys. Postpartum (PP) women and dams that had PE have an increased risk of developing HTN and chronic kidney disease (CKD) later in life. However, mechanisms linking PE to the long-term development of HTN and CKD are unknown. One aspect that may contribute to renal injury in PP PE women and dams is oxidative stress. Elevated concentrations of oxidative stress have been shown to augment the abundance and activity of renal transporters to increase sodium (Na+) reabsorption and blood volume. These alterations in renal transporters can consequently facilitate HTN. We hypothesize that at 6 weeks PP (~3 human years), PE dams will display oxidative stress, renal Na+ transporter abundance, and elevated blood pressure (BP). Pregnant Sprague Dawley rats were assigned to two groups: normal (CON) and PE dams. On gestational day 14, the reduced uterine perfusion pressure surgery was performed to generate a model of PE. Dams gave birth naturally and weaned for 3 weeks. After 6 weeks PP, BP was measured via carotid catheterization, and kidneys were removed and sectioned. Western blots were used to quantify renal Na+ transporters: Na+ K+ 2Cl-transporter (NKCC2) in the kidney medulla (KM) and epithelial Na+ channel (ENaC) in both the kidney cortex (KC) and KM. Oxidative stress was evaluated by heat shock protein 1 (HSP-1), copper zinc superoxide dismutase (CuZnSOD), and manganese superoxide dismutase (MnSOD) via Western blots. Hydrogen peroxide (H2O2) and antioxidant capacity concentrations were assessed via colorimetric assays. PP PE dams had increased BP (126.3±6.18vs105.7±3.74 mmHg, p<0.05) at 6 weeks after birth. KC HSP- 1, H2O2, MnSOD, and antioxidant capacity were unchanged between groups. However, KC CuZnSOD protein abundance was decreased in PP PE dams (69.51±11.64vs100±5.73 IU/Protein/Control%, p<0.05). In the KM, HSP-1 abundance (113.7±3.3vs100±5.07 IU/Protein/Control%, p=0.06) and H2O2 concentrations (1.97±0.11vs1.31± 0.38 nM H2O2/mg Protein, p=0.08) were elevated in PP PE dams. MnSOD, CuZnSOD, and antioxidant capacity were unchanged between groups in the KM. No changes occurred in KC and KM ENaC protein abundance. However, NKCC2 protein abundance was elevated by ~50% in PP PE dams (151.71±22.17vs100±5.59 IU/Protein/Control%, p=0.06). In summary, BP, oxidative stress, and NKCC2 were elevated in PP PE dams at 6 weeks. The presence of oxidative stress in the KM may lead to increased NKCC2 abundance. However, more studies are warranted to make this conclusion. NKCC2 elevation may result in increased Na+ and water reabsorption, leading to an increase in BP. Future studies will assess renal oxidative stress regulation of Na+ transporters in PP PE dams and determine the timeline PP in which changes in oxidative stress, Na+ transporters, and BP occur. This study is clinically relevant, because it indicates oxidative stress and NKCC2 in the KM, separately or together, may have a formative role in the pathogenesis of HTN and CKD in PP PE women later in life.Item LIMITED RESOURCES EARLY IN DEVELOPMENT ARE ASSOCIATED WITH HYPERTENSION LATER IN LIFE VIA CEREBRAL PROINFLAMMATORY CYTOKINE IL-17(2024-03-21) Burkes, Allison; Castillo, Angie; Smith, Savanna; Smith, Jonna; Jones, Kylie; Cunningham, MarkBACKGROUND: Poverty is pervasive and impacts nearly 17%of children in the U.S. While it is established that adverse childhood experiences (ACEs)such as poverty are associated with hypertension later in life, the mechanisms of this trend remain unclear. The limited bed and nesting (LBN) model simulates poverty in rodents by reducing available bedding by 80% for dams to create a nest for their offspring. The impact of inflammation on hypertension across the lifespan is unknown and is the focus of this study. Specifically, we evaluated 4-week and 17-week offspring to investigate the relationship of early-life stress and inflammation. METHOD: Timed pregnant Sprague Dawley dams gave birth naturally and their pups were weaned for 3 weeks. Postnatal days 2-9, dams and their pups were exposed to normal bedding (CON) or the LBN treatment. Offspring were divided by age, sex, and experimental status:at4weeks {LBN males (n=6), LBN females (n=3), CON males (n=5), CON females(n=5)} and at 17 weeks{ LBN males (n=5), LBN females (n=5), CON males (n=6) and CON females (n=6)}. On day 10, all rats were returned to normal bedding. At 4/17 weeks, offspring brain samples were harvested and analyzed via colorimetric assay to assess proinflammatory cytokineinterleukin-17 (IL-17). Carotid catheterizations were performed on 17-weekoffspring and blood pressures were measured. RESULTS: At 4 weeks, we found a trending difference between combined male and female LBN and CON groups(495.1 ±83.3 vs 291.1 ±64.8pg/mL/mg Protein; p=0.07,ns). Male offspring with LBN treatment had a significant increase in IL-17 concentration compared to CON males (964.3 ±108.5 vs 424.0 ±86.1; p<0.01). At 17 weeks, there was no longer a trending difference between LBN and CON groups with sexes combined, but there was a significant difference between male and female offspring with LBN and CON groups combined (7761 ±1005 vs 4511 ±901pg/mL/mg Protein; p<0.05). Although not significant, LBN Males had an upward trend of IL-17 compared to CON Males, while females showed no differences. Additionally, LBN Males had an upward trend in IL-17 compared to females exposed to LBN (9294 ±1279 vs 4198 ±1595pg/mL/mg Protein; p=0.08,ns). Blood pressure readings at sixteen weeks revealed LBN males have a significant increase in BP compared to CON males(128.17±3.93 vs 110.72±3.93 mmHg, p<0.05)and females showed no change(ns). CONCLUSION: Elevated cerebral IL-17 in male offspring suggests an increased inflammatory response to LBN treatment compared to females who showed no change. Accordingly, male sex may be a risk factor for high inflammatory responses to stress, like poverty. The differences in cytokine expression between males and females suggests that IL-17 contributes to hypertension experienced by LBN males later in life. More research is needed to understand the timeline of physiologic responses to ACEs and to investigate opportunities to intercept the inflammation response described in this study and literature. Future research should continue to develop an understanding of sex impacts on ACEs-related stress to refine treatment recommendations in the future.Item A new rodent model of preeclampsia: Pregnant daughters from hypertensive placental ischemic moms have hypertension(2023) Smith, Jonna; Powell, Madison; Cromartie, Whitney; Smith, Savanna; Jones, Kylie; Castillo, Angie; Wiemann, Natalia; McCafferty, Adair; Owen, Malissa; Srivastava, Prakriti; Cunningham, MarkPurpose: Studies show that daughters from hypertensive pregnancies are twice as likely to have preeclampsia (PE), pregnancy-induced hypertension (HTN) in comparison to women born from a normal pregnancy. PE affects ~5-10% of all births in the USA and is the leading cause of intrauterine growth restriction (IUGR). PE is associated with oxidative stress (OS) and cerebral damage. The causes of PE are unknown but is influenced by genetic and environmental conditions. Studies show that pregnancies involving placental insufficiency and HTN create an adverse environment that can affect the IUGR baby’s developmental programming and pregnancy outcomes. This study aims to characterize the pregnancy of IUGR rat offspring from hypertensive placental ischemic moms. We hypothesize female rats born from pregnant hypertensive placental ischemic moms will have elevated blood pressure (BP) and OS. Methods: Pregnant Sprague Dawley moms are divided into 2 groups: normal pregnant (NP) and the reduced uterine perfusion pressure (RUPP) hypertensive placental ischemic rats. On day 14 of pregnancy, the RUPP surgery is performed to generate PE. All dams (NP and RUPP) give birth naturally and weaned for 3 weeks. Offspring were then separated by sex and mother’s pregnancy status. ~10 weeks later, offspring were mated according to 4 groups: ♀NP x ♂NP (CON Preg, n=3), ♀NP x ♂RUPP (n=2), ♀RUPP x ♂NP (IUGR Preg, n=5), ♀RUPP x ♂RUPP (n=4). On day 19 of offspring pregnancy, BP was measured via carotid catheterization and the blood and brains were collected for analyses. Results: IUGR Preg rats have elevated BP (116 ± 4.17 vs 100.6 ± 2.54 mmHg, p<0.02) and 8-isoprostanes (439.2 ± 13.61 vs 381.3 ± 26.10 g, ns), decreased circulating antioxidant capacity (AC) (0.33 ± 0.01 vs. 0.37 ± 0.01 mM Trolox/mg protein, p<0.01), and reduced body weight (330.1 ± 5.24 vs 350.3 ± 10.82 g, ns) compared to CON Preg rats. IUGR Preg rats have larger brains, suggesting brain swelling (5.38 ± 0.10 vs 4.95 ± 0.19 g/1000g BW, p<0.04). HSP-1 (186.1 ± 28.14 vs 100.0 ± 6.36 %HSP-1/protein/CON, p<0.04) and H2O2 (25.76 ± 2.95 vs 15.81 ± 4.56 μM/mg protein, ns), markers of ROS, are increased in the brains of IUGR Preg vs. CON Preg rats. Cerebral AC was slightly reduced (260.0 ± 33.14 vs 292.3 ± 13.91 uM Trolox/mg protein) and MnSOD (antioxidant) amounts were decreased (87.96 ± 3.43 vs 100.0 ± 2.84 %MnSOD/protein/CON, p<0.63). Conclusion: IUGR Preg rats have increased systemic and cerebral OS, as well as larger brain sizes which may lead to cerebral damage. In summary, pregnant daughters from hypertensive placental ischemic moms show symptoms of a preeclamptic-like phenotype, thus creating a new model of PE. Future studies will determine the role of maternal PE status and OS in the development of HTN in pregnant IUGR offspring.Item Postpartum preeclamptic rats have hypertension and elevated cerebral oxidative stress(2023) Smith, Savanna; Smith, Jonna; Jones, Kylie; Castillo, Angie; McCafferty, Adair; Wiemann, Natalia; Owen, Malissa; Srivastava, Prakriti; Cunningham, MarkBackground: Postpartum (PP) preeclamptic (PE) women have an increased risk for developing hypertension (HTN), cerebrovascular diseases, and chronic kidney diseases later in life. The timing and mechanisms that contribute to a rise in blood pressure (BP), cerebrovascular and kidney dysfunction in PP PE women is unknown and the focus of this study. Previous studies in our lab (PMID: 34727994) indicate PP PE rats at 10 weeks have HTN and decreased antioxidant capacity (AC). Our current study examines BP and oxidative stress (OS) in PP PE rats at an earlier time point, 6 weeks (PP6). Understanding changes in cerebral and renal OS may reveal the pathophysiology of HTN, cerebrovascular, and renal disease development in PP PE women. We hypothesize that BP, renal, and cerebral OS will increase in PP6 PE rats. Methods: Pregnant Sprague Dawley rats were divided into 2 groups: control (CON) normal pregnant rats, and PE rats, derived from the surgically induced placental ischemic (reduced uterine perfusion pressure) model of PE. All rats gave birth and weaned for 3 weeks. At PP6, BP was measured via carotid catheterization. Brain and kidney tissues were collected to measure OS (HSP-1, Cu/ZnSOD, and MnSOD proteins and AC) through colorimetric assays and western blots. Results: PP6 PE vs CON rats, BP was elevated (128±6 vs 106±4mmHg, p<0.05) and AC was decreased in systemic circulation (28.5±5.1 vs 36.9±4.5mM Trolox/mg protein, ns). In the brain, both HSP-1 and Cu/ZnSOD were unchanged between PP6 PE and CON rats, while the levels of MnSOD (88.9±2.0 vs 100±2.5 IU/protein/CON %, p<0.05) and AC were decreased (619.1±179.2 vs 850.2±50.3 mM Trolox/mg protein, ns) in PP6 PE vs CON rats. In the kidney, HSP-1 decreased (88.7±3.0 vs 100±4.0 IU/protein/CON %, ns) in PP6 PE vs CON, while Cu/ZnSOD levels remained unchanged. However, kidney MnSOD levels significantly increased (124.3±8.0 vs 100±2.7 IU/protein/CON %, p<0.05) alongside an increase in AC (791.0±165.4 vs 587.0±64.5mM Trolox/mg protein, ns) in PP6 PE vs CON rats. Conclusion: PP6 PE rats have HTN and increased cerebral OS. Despite changes in the brain, kidneys appear to be protected from OS due to a decrease in a reactive OS protein (HSP-1) and increases in antioxidant capacity and protein (MnSOD). Future studies will determine the relationship between brain OS, HTN, and cerebral damage/dysfunction to PP PE rats. Furthermore, future studies will be designed to elucidate the protective mechanisms of the kidney in PP6 PE rats. Findings of this study are clinically relevant and could be used to improve the maternal health of women after PE pregnancies. In addition, therapy designed to target organ specific OS may be helpful in preventing HTN, cerebrovascular, and chronic kidney diseases later in life for women who have experienced PE.Item Renal Oxidative Stress May Explain Sex Differences in Blood Pressure in Adult Offspring Exposed to an Impoverished Environment(2024-03-21) Castillo, Angie; Smith, Savanna; Smith, Jonna; Jones, Kylie; Powell, MadisonNearly 40 million people experience poverty in the U.S. Poverty is linked to adverse childhood experiences (ACEs) which affects ~64% of adults in the U.S. Previous studies indicate that those who experience ACEs are at a higher risk of developing hypertension (HTN) and cardiovascular diseases (CVDs) later in life, with a greater severity and earlier onset in males. The mechanisms behind the ACEs-attributed development of sex differences in HTN later in life is unknown. One plausible mechanism for this sex difference is increased oxidative stress. One rodent model to mimic poverty as an ACE is the limited bed and nesting (LBN) model. This model simulates an impoverished and low resource environment, as observed in poverty, in which the nesting material during weaning is reduced. We hypothesize that male offspring exposed to LBN will have elevated blood pressure and oxidative stress, while females will have no change in blood pressure and reduced oxidative stress. Pregnant Sprague Dawley rats gave birth naturally and weaned their offspring for 3 weeks. During the weaning period, on Days 2-9, the dams and their respective pups were divided into 2 groups: LBN and control (CON). After LBN treatment, all rats received normal bedding. After weaning, offspring were divided by sex and experimental status: LBN male (n=5), LBN female (n=5), CON male (n=6), and CON female (n=6). At 16-17 weeks, mean arterial pressure (MAP) was measured via carotid catheterization and the kidneys, brain, heart, and plasma were collected to measure antioxidant capacity (AC) via colorimetric biochemical assays. LBN males had a significant increase (18 mmHg) in MAP compared to CON males (128.17±3.93 vs 110.72± 3.93 mmHg, P>0.001), while female LBN and CON rodents displayed no differences. In males, there were no changes in antioxidant capacity in the brain, heart, and plasma. However, there was a significant 2-fold decrease in renal antioxidant capacity (233±14.0 vs 442±12.3 mM Trolox/mg protein, p>0.0001). In females, there were no changes in the kidneys, heart, and brain antioxidant capacity. Conversely, LBN females showed a trending increase in plasma antioxidant capacity compared to CON (3.33±0.16 vs 2.53±0.35 mM Trolox/mg protein, p=0.053). Males exposed to an impoverished environment during weaning have elevated blood pressure, while females do not. This difference in blood pressure may be explained by decreases in renal AC in males only. On the other hand, females may be protected from elevated blood pressure because they experience a slight increase in systemic AC. Future studies will examine the role of antioxidants in blood pressure regulation in the kidneys. This is clinically relevant because ACEs affect a large percentage of the American population with ~17% of adults experiencing 4 or more ACEs, with minorities at a greater risk. Understanding the mechanisms on how ACEs contribute to HTN may alleviate some of the racial and ethnic disparities for people with HTN and CVDs. Perhaps, organ and sex-specific antioxidant therapies may prevent or reduce the development of HTN in adults that were exposed to ACEs, like poverty during childhood.